Aim To investigate the effects of osthol on bone marrow stromal stem cells in vitro under the con-ditions of the ability to differentiate into osteoblasts and the case of proliferation. Methods The rat bone marrow sample was obtained,and the all bone marrow cell culture methods were used to separate and collect the stratum of mononuclear cells. The cells were cultured in DMEM containing 10% fetal bovine serum. Three days later the culture medium was changed for the first time. Nine days later,serial subcultivation proceeded. The final concentration of osthol was 1 ? 10 -4,1 ? 10 -5,1 ? 10 -6,1 ? 10 -7 mol?L -1 respectively. MTT method was adopted in proliferation analysis. Under the induced condition,the Alkaline phosphatase activity, calcium salt sediment yield and osteocalcin were meas-ured on 4th,8th,12th,16th day. On the fifteenth day,his-tochemistry dyeing proceeded for calcified tubercle. Total RNA was isolated and the gene expression of bFGF, IGF-1,Osterix and Runx-2 was investigated by RT-Real Time PCR. Results The BMSCs proliferation was refrained by osthol dose-dependently. But it evidently led to osteogenesis. The ALP activity,calcium salt sediment yield and osteocalcin were raised,and calcified tubercle amount was increased. Besides,it also could enhance the mRNA level of bFGF,IGF-1,Osterix and Runx-2. Conclusion The osthol with final concentra-tion of 1 ? 10 -5 mol?L -1 can markedly promote BM-SCs differentiation to osteogenesis. which proves osthol is an active constituent of the traditional Chinese medi-cine Common Cnidium Fruit.

This study is to investigate the effects on the expression of iNOS and production of NO in the osteogenic differentiation process of rat bone marrow stromal cells (rBMSCs) by icariside II. rBMSCs were cultured by adherence screening method. When the culture dishes were covered with 80% cells, the osteogenic induced cultures were adopted. Icariside II was supplemented into the culture at 1 x 10(-5) mol x L(-1). The activity of iNOS, content of NO and osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-FALP and mineralized bone nodules were compared among the icariside II-supplemented group, L-NMAE group, icariside II + L-NAME group and the control. Total RNA was isolated and the gene expression of iNOS, Osterix and Runx-2 was investigated by real-time PCR. Total protein was also isolated and the secretion of iNOS and collagen I was examined by Western blotting. Icariside II can significantly improved ALP activity, CFU-FALP amount and mineralized nodules. Besides, the mRNA level of factors related to the osteogenic differentiation includes Osterix and Runx-2 also enhanced. The secretion of collagen I also promoted significantly. But all of these effects can be inhibited by L-NAME which can specifically inhibit the activity of iNOS. Icariside II enhances the osteogenic differentiation of rBMSCs significantly, but if the activity of iNOS was blocked by L-NAME, the osteogenic differentiation markers decrease accompanied with iNOS and NO decrease, suggesting that icariside II stimulates the osteogenic differentiation via enhancing the activity of iNOS and promoting the generation of NO.

OBJECTIVETo investigate the effects of icariin on the osteogenic differentiation of rat bone marrow stromal cells (rBMSCs).

METHODrBMSCs were cultured by adherence screening method. Icariin was supplemented into the culture at 1 x 10(-5) mol x L(-1). The osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-F(ALP) and mineralized bone modulus were compared between the icariin-supplemented group and the control group. Total RNA was isolated and the gene expression of bFGF, IGF-1, Osterix(OSX) and Runx-2 was investigated by RT Real-time PCR.

RESULTIcariin significantly improved ALP activity, CFU-F(ALP) amounts and mineralized modulus. It also can enhance the mRNA level of bFGF, IGF-1, Osterix and Runx-2.

CONCLUSIONIcariin enhances the osteogenic differentiation of rBMSCs significantly, which suggested that icariin has the potentiality to be a new drug of anti-osteoporosis or fracture healing.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Male ; Osteogenesis ; drug effects ; Rats ; Rats, Wistar ; Stromal Cells ; cytology ; drug effects ; metabolism

OBJECTIVETo investigated the effects of isopsoralen on bone marrow stromal stem cells (BMSCs) differentiate and proliferate in vitro.

METHODThe stratum of mononuclear cells were separated and collected from the rat bone marrow sample by the all bone marrow cell culture methods. The cells were cultured in DMEM contained 10% fetal bovine serum. The culture medium was changed after three days. Nine days later, cells were treatment by isopsoralen with the concentration 1 x 10(-5), 1 x 10(-4), 1 x10(-6), 1 x 10(-7) mol x L(-1). MTT method was used for the proliferated analyzing. Under the induced condition, the alkaline phosphatase (ALP) activity, calcium salt sediment yield and osteocalcin were measured at the 4, 8, 12, 16 d. At the fifteenth day, histochemistry dyeing for calcified tubercle and ALP was proceeded. Total RNA was isolated and the gene expression of bFGF, IGF-1, Osterix and Runx-2 were investigated by Real Time PCR.

RESULTThe BMSCs proliferation refrained by isopsoralen with dose dependent. But it significantly enhanced osteogenesis, which was represented by the promotion of the ALP activity, calcium salt sediment yield, osteocalcin, and calcified tubercle amount. Besides, it also enhanced the mRNA level of bFGF, IGF-1, Osterix and Runx-2.

CONCLUSIONThe isopsoralen with the concentration 1 x 10(-5) mol x L(-1) can promote BMSCs differentiation to osteoblasts. It demonstrated the isopsoralen can prevent antiosteoporotic, which is an active part of the traditional Chinese medicine psoralea corylifolia.

Alkaline Phosphatase ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; genetics ; Cell Proliferation ; drug effects ; Core Binding Factor Alpha 1 Subunit ; genetics ; Fibroblast Growth Factors ; genetics ; Furocoumarins ; pharmacology ; Gene Expression Regulation ; drug effects ; Insulin-Like Growth Factor I ; genetics ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteocalcin ; metabolism ; Osteogenesis ; drug effects ; RNA, Messenger ; genetics ; Rats ; Stromal Cells ; cytology ; drug effects ; metabolism ; Transcription Factors ; genetics

Clinical studies have confirmed that cryoablation is a safe and effective treatment for lung cancer. Cryoablation has been clinically used in the treatment of various types of lung cancer,and has achieved good therapeutic effects. Some of the complications of cryoablation can be alleviated after symptomatic treat-ment. However,cryoablation still needs further research and exploration in clinical applications.